A kampo preparation is a pharmaceutical prepared by blending crude drugs, in which all the active ingredients are not always specifically identified. Furthermore, a single active ingredient alone does not always exhibit its effect, and some active ingredients may compositely act with each other. For securing its quality, it is said that an assay method capable of totally evaluating the whole kampo preparation is necessary (Patent Document 1, Patent Document 2).
The assay method includes a method of total evaluation by assaying the individual ingredients, and a bioassay of evaluating the physiological activity by using a biological material. The bioassay includes an in-vivo test and an in-vitro test, and the in-vivo test system has many limitations regarding the test facilities, test animals, processing capability, and the like, and there were some difficulties in applying the in-vivo test to the quality evaluation of kampo preparations.
On the other hand, the in-vitro test system does not require any special facilities and gives stable test results in a short period of time. For this reason, it is desired to establish a bioassay method with this system. In fact, for myostatin, a bioassay method is reported (Patent Document 3). However, for a kampo preparation that comprises a combination of crude drugs each having plural active ingredients by themselves, a suitable bioassay system could not always be found out, and the establishment of the bioassay system is desired.
For example, yokukansan, a type of kampo preparation, generally has the composition shown below. A suitable bioassay system is not yet found out also for yokukansan. For securing higher quality for yokukansan, the development of the bioassay system for yokukansan is desired.
TABLE 1IngredientsAmountJP Atractylodes Lancea Rhizome4.0 gJP Poria Sclerotium4.0 gJP Cnidium Rhizome3.0 gJP Japanese Angelica Root3.0 gJP Bupleurum Root2.0 gJP Glycryrrhiza1.5 gJP Uncaria Hook3.0 g    Patent Document 1: JP-T 2000-512621    Patent Document 2: JP-T 2001-521876    Patent Document 3: JP-T 2005-520486    Non-Patent Document 1: “Planta Med.” 2004 May; 70(5): 427-31 (Hong H, Liu G Q. Scutellarin attenuates oxidative glutamate toxicity in PC12 cells)    Non-Patent Document 2: “J. Ethnopharmacol.” 2005 Apr. 8; 98(1-2): 89-94 (Yu D, Duan Y, Bao Y, Wei C, An L. Isoflavonoids from Astragalus mongholicus protect PC12 cells from toxicity induced by L-glutamate)    Non-Patent Document 3: “Clip. Exp. Pharmacol. Physiol.” 2004 August; 31(8): 530-6 (Lee J H, Song D K, Jung C H, Shin D H, Park J, Kwon T K, Jang B C, Mun K C, Kim S P, Suh S I, Bae J H, (−)-Epigallocatechin gallate attenuates glutamate-induced cytotoxicity via intracellular Ca modulation in C12 cells)